Daphnia is a good model organism for studying factors affecting dispersal and patterns of genetic diversity. Within this genus, the Daphnia pulex species complex includes lineages from North America and Europe, with some considered invaders in various continents, although their colonization history is poorly known. We used mitochondrial DNA and microsatellite markers to identify the D. pulex complex lineages in Sierra Nevada, determine their reproductive mode and reconstruct their genetic history (over the past ~25 to 65 years). We present the first recording of North American (NA) D. cf. pulex in a European high-mountain lake, showing its arrival ~65 years ago in lake Borreguil without temporal changes in its genetic structure. European (Eu) D. cf. pulicaria is the only lineage present in other Sierra Nevada lakes and also showed no genetic change over time. The results for both species are congruent with obligate parthenogenetic reproduction mode. Moreover, water mineralization may influence the clonal distribution of the D. pulex complex in Sierra Nevada, without ruling out dispersal limitation and/or founder effects. Although NA D. cf. pulex had not spread to other Sierra Nevada lakes, it could threaten Eu D. cf. pulicaria in Sierra Nevada and other European alpine lakes.
The biodiversity of Mediterranean freshwater bodies is among the most threatened worldwide; therefore, its accurate estimation is an urgent issue. However, traditional methods are likely to underestimate freshwater zooplankton biodiversity due to its high species seasonality and cryptic diversity. We test the value of applying DNA barcoding to diapausing egg banks, in combination with the creation of a reference collection of DNA barcodes using adult individual samples, to characterize rotifer communities. We use monogonont rotifers from two lakes in Doñana National Park and one from Ruidera Natural Park in Spain as models to create a reference collection of DNA barcodes for taxonomically diagnosed adult individuals sampled from the water column, to compare with the sequences obtained from individual eggs from the diapausing egg banks. We apply two different approaches to carry out DNA taxonomy analyses, the generalized mixed Yule coalescent method (GMYC) and the Automatic Barcode Gap Discovery (ABGD), to the obtained sequences and to publicly available rotifer sequences. We obtained a total of 210 new rotifer COI sequences from all three locations (151 diapausing eggs and 59 adults). Both GMYC and ABGD generated the same 35 operational taxonomic units (OTUs), revealing four potential cryptic species. Most sequences obtained from diapausing eggs (85%) clustered with sequences obtained from morphologically diagnosed adults. Our approach, based on a single sediment sample, retrieved estimates of rotifer biodiversity higher than or similar to those of previous studies based on a number of seasonal samples. This study shows that DNA barcoding of diapausing egg banks is an effective aid to characterize rotifer diversity in Mediterranean freshwater bodies.
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